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Biomedical subjects

N Shindo

Publications and source records attributed to N Shindo.

At least 37 records · Page 2Linked to original sources
PubMed1996

Optical resolution of phenylthiohydantoin-amino acids by capillary electrophoresis and identification of the phenylthiohydantoin-D-amino acid residue of [D-Ala2]-methionine enkephalin.

This is an initial report to propose a protein sequence analysis system with DL differentiation using capillary electrophoresis (CE). This system consists of a protein sequencer and a CE system. After fractionation of phenyl-thiohydantoin (PTH)-amino acids using a protein sequencer, optical resolution for each PTH-amino acid is performed by CE using some chiral selectors such as digitonin, beta-escin and others. As a model peptide, [D-Ala2]-methionine enkephalin (L-Tyr-D-Ala-Gly-L-Phe-L-Met), was used and the sequence with DL differentiation was determined, with the exception of the fourth amino acid, L-Phe, using our proposed system.

Amino Acids↗
PubMed1996

Studies on the mechanism of early onset macular degeneration in cynomolgus monkeys. II. Suppression of metallothionein synthesis in the retina in oxidative stress.

Initial investigations done in this laboratory detected increased albumin and decreased glyceraldehyde 3-phosphate dehydrogenase concentrations in the retina of an animal model manifesting early onset macular degeneration. Both glyceraldehyde 3-phosphate dehydrogenase and albumin are markers of oxidative stress in cells. In this study, we used the same animal model to study further biochemical and physiological processes which may be involved in the pathogenesis of early onset macular degeneration in monkeys. We detected 60% lower catalase and glutathione peroxidase activities in the affected retinas suggesting lower antioxidant activities and oxidative stress. One of the consequences of oxidative stress is the production of metallothionein, a low molecular weight protein also induced by high concentrations of heavy metals such as zinc. Metallothionein was detected by RT-PCR in these monkey retinas. However initial quantitative PCR studies on this protein showed that the synthesis of metallothionein in affected retinas appears to be less than in normal controls. The affected retinas also showed a fourfold lower zinc concentration compared with the normal controls. No significant difference, however, could be detected in the zinc concentrations in plasma samples. Since induction of metallothionein synthesis is mediated by transcription factors which require heavy metals such as zinc for binding to specific sites in the DNA, the lowered zinc concentration may, thus, correlate with the lowered metallothionein expression. And since metallothionein is suggested to function as a free radical scavenger, the lowered metallothionein synthesis may consequently contribute to increased peroxidation reactions in the affected retinas. It appears therefore, that oxidative stress and the decreased metallothionein synthesis may be involved in the pathogenesis of early onset macular degeneration in this animal model.

Amino Acid Sequence↗
PubMed1996

Electron microscopic study of chronic eosinophilic pneumonia.

Two cases of chronic eosinophilic pneumonia were examined electron microscopically to study the role of eosinophil granulocytes. Eosinophils, together with macrophages and lymphocytes, were observed to have infiltrated prominently in the lung tissues of the two cases. Degeneration and necrosis of pneumocytes were observed, and denuded basement membranes and destroyed alveolar structures were occasionally found. In the alveolar septa, swelling of the endothelial cells of the blood capillaries, edema of the stroma and fibrin deposition were observed. Eosinophils, released eosinophil granules and macrophages phagocytosing eosinophil granules were found frequently near the degenerated and necrotic alveolar tissues. These findings suggest that the tissue injuries were induced by the cytotoxic effects of the eosinophil granules.

Adult↗
PubMed1996

Sepsis associated with hematological malignancies: prophylaxis of Pseudomonas aeruginosa sepsis.

Underlying diseases, pathogenic bacteria, clinical background and outcome were studied during 91 febrile episodes complicated by sepsis in 55 patients with hematological malignancies, who had been admitted to our hospital (Jikei University Kashiwa Hospital) between January 1990 and December 1994. Particularly in patients with P. aeruginosa sepsis, we compared the prophylactic effect of ciprofloxacin (CPFX) alone with that of the combination of polymyxin B (PL-B) plus kanamycin (KM). The major underlying diseases were acute myelocytic leukemia and malignant lymphoma, followed by myelodysplastic syndrome, acute lymphocytic leukemia and chronic myelocytic leukemia. Nearly two-thirds of the pathogenic microorganisms isolated were gram-positive bacteria (including coagulase-negative staphylococci and Staphylococcus aureus); approximately one-quarter were gram-negative bacteria (such as Pseudomonas aeruginosa), and the remainder were fungi. These microorganisms usually induced sepsis when granulocyte counts were decreased. Sepsis was a direct cause of death in about 60% of the patients and P. aeruginosa sepsis had the worst outcome. Oral administration of CPFX was more effective than PL-B plus KM in preventing P. aeruginosa sepsis. The difference in effectiveness might depend on the absorption profile of the drugs.

Adolescent↗
PubMed1996

[Two cases of gram-positive sepsis successfully treated with vancomycin in combination with imipenem or cilastatin].

Against gram-positive sepsis, vancomycin (VCM) was administered in combination with imipenem or cilastatin (IPM/CS). Its excellent efficacy was confirmed in 2 cases, one affected with methicillin resistant Staphylococcus aureus and another with Gemella morbillorum. By calculating the FIC index according to a checkerboard technique, the in vitro synergistic effect was also demonstrated. At the present state multi-drug resistant gram-positive infections prevailed, the combination of VCM with IPM/CS can be expected as an effective measure for treating these diseases.

Adolescent↗
PubMed1994

Proteinuria induced by anti-dipeptidyl peptidase IV (gp108); role of circulating and glomerular antigen.

Massive proteinuria is induced in rats by administration of rabbit antibody to dipeptidyl peptidase IV (DPPIV, gp108), a glycoprotein present on glomerular cell membranes and in serum. This study was undertaken to know which antigen, glomerular or serum DPPIV, is responsible for forming immune complex in glomeruli and development of proteinuria. An i.p. injection of the antibody resulted in a rapid decrease of serum DPPIV and a gradual increase of rabbit IgG deposited along glomerular capillary wall for 4-8 h. Abnormal proteinuria appeared within 8 h, peaked on day 2 (> 200 mg/24 h) and then declined. An increase of urinary protein and glomerular deposition of IgG also occurred, when the antibody was injected into serum DPPIV-depleted rats that had received preinjection of anti-DPPIV antibody. These results suggest that proteinuria is induced by direct binding of anti-DPPIV antibody to the membrane antigen of glomerular cells.

Animals↗
PubMed1993

Molecular cloning and sequence analysis of a segment from Andean potato mottle virus B RNA encoding the putative RNA polymerase.

Andean potato mottle virus (APMV), an endemic South American comovirus, has a bipartite genome consisting of two plus-strand RNA molecules (M and B RNA). We have cloned the 3' half of the B RNA and identified the complete sequence of the putative APMV RNA polymerase. The RNA polymerase gene is part of a large polyprotein-encoding open reading frame. The putative, mature RNA polymerase, as deduced by comparison with the related cowpea mosaic virus (CPMV), type member of the comovirus group, is 703 amino acids long and shows a large degree of similarity with CPMV and other RNA polymerases.

Amino Acid Sequence↗
PubMed1993

Nucleotide sequence analysis of an Andean potato mottle virus middle component RNA cDNA clone: comparisons of the encoded proteins with those of other comoviruses.

Andean potato mottle virus (APMV) is a comovirus whose genomic structure consists of two plus-strand RNA molecules (M- and B-RNA). Here we report the nucleotide sequence analysis of an APMV M-RNA cDNA clone with 3,669 nucleotide (nt) residues, exclusive of the polyadenylate at the 3' end, covering approximately 99% of the APMV M-RNA. The first initiation codon in register translates from nt 194 to 3185 a polyprotein of 997 amino acid (aa) residues. A second initiation codon in register, beginning at nt position 416, translates a polyprotein of 923 aa. The cleavage sites used in the processing of polyprotein were identified in the long open reading frame by N-terminal microsequencing of the large coat protein (LCP) and the small coat protein (SCP). These dipeptide cleavage sites are Q/M for the LCP and Q/F for the SCP. In a comparison of the deduced APMV polyprotein aa sequence with those of four other comoviruses, the coding regions for the putative movement protein, LCP and SCP, were found similar in length in all five species. Multiple alignment of the M-RNA sequences for each of the three genes from the five comoviruses revealed different degrees of homology. APMV was always the least homologous of the five comoviruses, showing significant aa substitutions in positions where the other comoviruses have identical residue or conservative substitutions.

Amino Acid Sequence↗
PubMed1990

Capillary gas chromatography/negative ion chemical ionization mass spectrometry for the quantification of bile acids including 1 beta-hydroxylated and unsaturated bile acids in serum and urine.

12 bile acids, including 1 beta-hydroxylated and unsaturated bile acids, have been quantified by capillary gas chromatography/negative ion chemical ionization mass spectrometry, using the trimethylsilyl(TMS) ether derivatives of bile acid pentafluorobenzyl(PFB) esters. The analysis time is 12 min and the minimum measurable amount is 100 fg for each bile acid. Bile acids in 200 microL of serum and 50 microL of urine from healthy human adults were measured. These small sample sizes enhance the practicality of using this method as a screening test for bile acids in the serum and urine of human infants, where small sample size is a major problem.

Adult↗
PubMed1990

Identification of 17-hydroxyprogesterone and other steroid hormones in saliva from a normal child and patients with congenital adrenal hyperplasia by plasmaspray liquid chromatography/mass spectrometry.

A very sensitive, selective and simultaneous method for the analysis of salivary steroid hormones was examined by discharge-assisted thermospray liquid chromatography mass spectrometry (plasmaspray LC/MS). Plasmaspray LC/MS gave [M + H]+ or [MH - H2O]+ as a predominant ion in most of the steroids in this work and in some cases fragment ions were also observed. Eight salivary steroid hormones, pregnenolone, 17-hydroxypregnenolone, progesterone, 17-hydroxyprogesterone, 11-deoxycortisol, cortisol, aldosterone, and testosterone were identified within 10 minutes using the selected ion monitoring technique in conjunction with plasmaspray LC/MS. Progesterone, 17-hydroxyprogesterone, cortisol, aldosterone and testosterone were detected in 1 mL of saliva from a normal child and two patients with congenital adrenal hyperplasia.

17-alpha-Hydroxyprogesterone↗
PubMed1990

Plasma osteocalcin in preparturient and postparturient cows: correlation with plasma 1,25-dihydroxyvitamin D, calcium, and inorganic phosphorus.

The purpose of the present study was to determine changes of plasma osteocalcin levels in periparturient cows and to examine the correlation of plasma osteocalcin level with that of 1,25-dihydroxyvitamin D, Ca, and inorganic P. Five Holstein-Friesian cows, aged 3 to 5 yr, were used from 5 d before to 15 d after calving. Concentration (mean +/- SE) of plasma osteocalcin decreased rapidly from d 1 prepartum, reached a low of 8.0 +/- 3.0 ng/ml at d 1 postpartum, and then recovered gradually to 17.8 +/- 3.8 ng/ml at 15 d after calving. In contrast, the concentration of plasma 1,25-dihydroxyvitamin D increased from calving to 3 d postpartum. Plasma concentrations of osteocalcin did not correlate significantly with those of 1,25-dihydroxyvitamin D, but it was significantly proportional to that of Ca and inorganic P. It is suggested that osteoblast function is depressed by a number of factors around the time of parturition.

Animals↗
PubMed1990

A case of GM1-gangliosidosis type I: glycosphingolipid profiles of urine and transformed lymphocytes and beta-D-galactosidase activities in peripheral lymphocytes, cultured skin fibroblasts and transformed lymphocytes.

A female infant with early-onset GM1-gangliosidosis type I was investigated. The lymphocytes, transformed lymphocytes and cultured skin fibroblasts of the patient were demonstrated to have severe beta-D-galactosidase deficiency. The beta-D-galactosidase activities of these cells from the patient's father and mother were at the lower limit of the normal range. The oligosaccharide accumulation in urine of the patient showed the typical type I GM1-gangliosidosis pattern, but no GM1 ganglioside was detected in the patient's urine or transformed lymphocytes. The clinical features were compatible with infantile GM1-gangliosidosis. The mixture of homogenates from the cultured fibroblasts or transformed lymphocytes of the patient and controls showed no complementation of beta-D-galactosidase activity against the controls.

Cells, Cultured↗
PubMed1989

Biochemical mechanisms of aminoglycoside cell toxicity. II. Accumulation of phospholipids during myeloid body formation and histological studies on myeloid bodies using twelve aminoglycoside antibiotics.

Cultured human skin fibroblasts take up aminoglycoside antibiotics into lysosomes to form myeloid bodies. Gentamicin (GM), one such antibiotic, was taken up until the cellular concentration reached an estimated 64 mM on the 3rd day when cells were incubated with 2 mM gentamicin. The rate of release of intracellular GM was high on the first day of incubation and gradually slowed down over the next 4 d. About 50% of the GM remained in the cells even on longer incubation in GM-free medium, suggesting it may irreversibly bind to cellular components. With myeloid body formation, the cellular phospholipid content increased 1.5 times. Bis(monoacyl-glyceryl)phosphate, which is known as a marker of lysosomal phospholipid, phosphatidylcholine and phosphatidylserine showed 250, 162, and 153% increases, respectively. Sphingomyelin was not accumulated, while lysosomal sphingomyelinase was dramatically inhibited. Of 12 different aminoglycoside antibiotics, paromomycin is the most prominent myeloid body-forming antibiotic. The myeloid body-formation is not directly correlated to human nephrotoxicity. On the other hand, the number of myeloid bodies is well correlated to the affinity to the brush border membrane, suggesting that such aminoglycoside antibiotics are taken up easily through cellular endocytosis. The cytotoxic effects of aminoglycoside antibiotics may be due to by their binding to cellular organelles other than lysosomes.

Anti-Bacterial Agents↗
PubMed1989

[A case of mixed müllerian tumor, usefulness of 67Ga-citrate scintigraphy].

Mixed Müllerian Tumor (M.M.T.) is a rare malignant neoplasm usually arising from uterine body in postmenoposal woman, and imaging diagnosis for the tumor has not been established. A 45-year-old female with pathologically confirmed M.M.T. was evaluated by 67Ga-citrate scintigraphy and CT on the basis of imaging diagnosis. 67Ga-citrate scintigraphy detected correctly not only primary lesions but also metastatic and recurrent-foci. The findings of 67Ga-citrate scintigraphy were confirmed to be correspond to autopsy findings. Generally, detectability of 67Ga-citrate scintigraphy for genito-urinary tract neoplasm was not appreciated, but in our case 67Ga-citrate scintigraphy was useful for detection of the primary lesion, metastatic and recurrent foci. The cellular components and grade of cell-differentiation of this tumor seemed to be responsible for good detection of the tumor. Thus, 67Ga-citrate scintigraphy was considered to be useful diagnostic imaging modality of M.M.T.

Female↗
PubMed1989

[Preliminary study of patients with vertigo and dizziness using 123I-IMP].

To study the application of cerebral blood flow scintigraphy with 123I-IMP for the evaluation of central dizziness, the difference in 123I-IMP accumulation between the right and left cerebellar hemispheres was determined as an asymmetry index (A.I.) in 8 normal subjects and compared with A.I. from patients with central dizziness. According to estimated lesions, 123I-IMP findings were compared with CT findings. The normal subjects had A.I. of 5.8 +/- 2.1%, whereas 36 patients with central dizziness had A.I. of 12.7 +/- 7.5%. On the basis of an A.I. of 8% or more as an abnormal finding on IMP scintigraphy, 11 of the 14 patients with estimated lesions of cerebellar hemispheres and 16 of the 20 with those of the whole cerebellum including the vermis were evaluated to have had abnormalities. Seven of 11 with estimated lesions of the brainstem and 4 of 5 with vertebrobasilar arterial insufficiency were evaluated to have had abnormalities. The findings of 123I-IMP cerebral blood flow scintigraphy were in good agreement with clinical symptoms in patients with central dizziness with estimated lesions of the cerebellar hemispheres, suggesting the possibility that this method is more useful than CT.

Adult↗